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通过聚合酶链反应和测序分析检测拉丁美洲反应性关节炎患者的细菌DNA。

Detection of bacterial DNA in Latin American patients with reactive arthritis by polymerase chain reaction and sequencing analysis.

作者信息

Cuchacovich Raquel, Japa Shankar, Huang Wen Qun, Calvo Armando, Vega Luis, Vargas Ruben Burgos, Singh Ranju, Flores Diana, Castro Ivette, Espinoza Luis R

机构信息

Department of Medicine, LSU Health Science, Center, New Orleans, Louisiana 70112, USA.

出版信息

J Rheumatol. 2002 Jul;29(7):1426-9.

Abstract

OBJECTIVE

Bacteria and/or their antigens are thought to play a role in the pathogenesis of reactive arthritis (ReA). Polymerase chain reaction (PCR) using the 16S ribosomal RNA-PCR method was used to identify bacterial DNA in synovial fluid (SF) and tissue (ST) in a well defined group of patients with chronic ReA. In addition, species found were identified by means of sequence analysis.

METHODS

We examined 15 ST and 5 SF samples of 15 patients with ReA, 5 ST samples of 5 patients with osteoarthritis (OA), and 8 SF from 8 patients with closed traumatic knee injuries using a nested PCR with universal 16S rRNA primers. In addition, a nested PCR was developed to detect DNA sequences of Salmonella sp. and Mycoplasma sp. Automated sequencing and comparative data analysis (GenBank) were also performed to identify the species.

RESULTS

Bacterial DNA was identified in 8 cases, 5 ST and 3 SF; Chlamydia trachomatis (n = 2), Pseudomonas sp. (n = 3), and Bacillus cereus (n = 2) were the most common microorganisms identified. A variety of microorganisms including Clostridium sp., Lactobacillus sp., Pseudomonas migulae, P. fluorescens, and P. putida, and Neisseria meningitidis serogroup B were also identified. In half of the cases (4/8) 2 to 3 bacterial antigens were identified simultaneously.

CONCLUSION

Bacterial DNA is present in the joints in patients with chronic ReA. A wide spectrum of bacteria including some not previously associated with ReA were identified. Further studies are needed to establish their exact role in the pathogenesis of ReA and related arthritides.

摘要

目的

细菌和/或其抗原被认为在反应性关节炎(ReA)的发病机制中起作用。采用16S核糖体RNA聚合酶链反应(PCR)方法,对一组明确诊断的慢性ReA患者的滑液(SF)和组织(ST)中的细菌DNA进行鉴定。此外,通过序列分析对所发现的菌种进行鉴定。

方法

我们使用通用16S rRNA引物进行巢式PCR,检测了15例ReA患者的15份ST样本和5份SF样本、5例骨关节炎(OA)患者的5份ST样本以及8例闭合性膝关节创伤患者的8份SF样本。此外,还开发了一种巢式PCR来检测沙门氏菌属和支原体属的DNA序列。同时进行自动测序和比较数据分析(GenBank)以鉴定菌种。

结果

在8例样本中鉴定出细菌DNA,其中5份ST样本和3份SF样本;沙眼衣原体(n = 2)、假单胞菌属(n = 3)和蜡样芽孢杆菌(n = 2)是最常见的鉴定出的微生物。还鉴定出了多种微生物,包括梭菌属、乳杆菌属、米古拉假单胞菌、荧光假单胞菌、恶臭假单胞菌以及B群脑膜炎奈瑟菌。在半数病例(4/8)中同时鉴定出2至3种细菌抗原。

结论

慢性ReA患者的关节中存在细菌DNA。鉴定出了多种细菌,包括一些先前未与ReA相关联的细菌。需要进一步研究以确定它们在ReA及相关关节炎发病机制中的确切作用。

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